Search Results (55,210)

High Mobility Group ^{Box 1} (HMGB1) is a central pro-inflammatory mediator released from damaged or stressed cells, where it activates receptors such as the Receptor for Advanced Glycation Endproducts (RAGE). Dendritic polyglycerol sulfate (dPGS), a hyperbranched polyanionic polymer, is known for its anti-inflammatory activity. In this study, we examined how dPGS modulates HMGB1-driven signaling in RAW 264.7 macrophages and human microglia. Recombinant human HMGB1 expressed in Escherichia coli (E. coli) was purified by nickel-nitrilotriacetic acid (Ni-NTA) and heparin chromatography. Proximity ligation assays (PLA) revealed that dPGS significantly disrupted HMGB1/RAGE interactions, particularly under lipopolysaccharide (LPS) stimulation, thereby reducing inflammatory signaling complex formation. This correlated with reduced activation of the nuclear factor kappa B (NF-κB) pathway, demonstrated by decreased nuclear translocation and transcriptional activity. Reverse transcription polymerase chain reaction (RT-PCR) and quantitative real-time PCR (RT-qPCR) showed that dPGS suppressed HMGB1- and LPS-induced transcription of tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6), monocyte chemoattractant protein-1 (MCP-1), cyclooxygenase-2 (COX-2), and inducible nitric oxide synthase (iNOS). Enzyme-linked immunosorbent assay (ELISA) and Griess assays confirmed reduced TNF-α secretion and nitric oxide production. Electron paramagnetic resonance (EPR) spectroscopy further showed that dPGS altered HMGB1/soluble RAGE (sRAGE) complex dynamics, providing mechanistic insight into its receptor-disruptive action. Full article

Salinity is a pivotal environmental factor that significantly influences the survival, growth, development, and reproduction of aquatic organisms. However, the characteristics of serum metabolites and their mechanistic roles in mediating the response of grass carp (Ctenopharyngodon idellus) to long-term salinity stress remain incompletely understood. Therefore, the present study exposed grass carp to different salinity levels (0, 4, and 8 g/L) for 60 days to evaluate the associated physiological alterations and metabolic responses. The results revealed that high salinity (8 g/L) significantly suppressed growth performance (p < 0.05), whereas low salinity (4 g/L) caused no significant reduction in growth or survival. Physiological analyses indicated that fish in the 8 g/L group exhibited markedly reduced levels of lactic acid and total protein, along with elevated concentrations of total cholesterol, triglycerides, glucose, and glutamic-oxaloacetic transaminase (p < 0.05). Serum ion homeostasis was also disrupted under high salinity, characterized by increased Ca²⁺, Na⁺, and Cl⁻ levels and decreased Mg²⁺ (p < 0.05). Furthermore, oxidative stress was evident in the high-salinity group through heightened activities of antioxidant enzymes (SOD, CAT, GPx), accumulation of oxidative damage markers (protein carbonyl, 8-OHdG) (p < 0.05). Metabolomic profiling identified 367 and 403 significantly altered metabolites in the 4 g/L and 8 g/L groups, respectively, primarily belonging to lipids and lipid-like molecules along with organic acids and derivatives. KEGG enrichment analysis revealed that these differential metabolites were chiefly involved in amino acid biosynthesis, glycerophospholipid metabolism, biosynthesis of unsaturated fatty acids, and glycine, serine, and threonine metabolism. Trend analysis further uncovered eight distinct expression patterns of metabolites across salinity gradients. These results provide novel insights into the metabolic adaptations of grass carp to salinity stress, demonstrating that high salinity induces oxidative stress, disrupts ion regulation, and drives extensive metabolic reprogramming. The study offers valuable theoretical support for improving salinity tolerance management in aquaculture and informs the selective breeding of salt-tolerant fish strains. Full article

Periodontitis (PD) is a multifactorial, progressive inflammatory disease affecting the teeth-supporting tissues, characterized by an imbalance of the oral microbiota and the presence of bacterial biofilms leading to host response. Nowadays, reliable biochemical markers for early and objective diagnosis, and for predicting disease progression, are still lacking. Our previous proteomic investigations revealed the significant overexpression of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) in periodontal pocket tissue, gingival crevicular fluid (GCF), and tooth-surface-collected material (TSCM) from PD patients in comparison to periodontally healthy controls, proposing it as a possible biomarker of PD. This study aimed to evaluate the expression of GAPDH in saliva, a more accessible, non-invasive, and clinically relevant oral sample. The whole saliva was analyzed by a preliminary mass spectrometry-based proteomic approach, identifying significantly increased levels of GAPDH also in salivary samples from periodontal-affected subjects. These data were further validated by enzyme-linked-immunosorbent assay (ELISA). Additionally, protein–protein interaction networks were generated through the Human Protein Atlas database, using different datasets (OpenCell, IntAct, and BioGRID). Bioinformatic analysis provided noteworthy GAPDH-associated networks potentially relevant to periodontal pathology. The scientific significance of this study lies in the detection of salivary GAPDH as a novel strategy to advance periodontal clinical diagnostics from the perspective of a non-invasive screening test. In correlation with other protein markers, salivary GAPDH could constitute a promising set of distinctive and predictive targets to enhance early diagnosis of PD, disease monitoring, and treatment planning in periodontology. Full article

Background/Objectives: The purpose of this study was to investigate the association between HER-2 expression and clinicopathological characteristics, biochemical recurrence (BCR) rate, and BCR-free survival in localized prostate cancer (PCa) patients after radical prostatectomy (RP). Methods: Between January 2018 and December 2019, 44 patients with pathologically confirmed localized PCa who underwent RP were included in this study. According to the expressed level of HER-2 protein, patients were divided into four cohorts: cohort-1 (HER-2 0), cohort-2 (HER-2 1+ or 2+), cohort-3 (HER-2 0 or 1+), and cohort-4 (HER-2 2+); the clinicopathological and clinical outcomes were analyzed and compared between cohort-1 and cohort-2, and cohort-3 and cohort-4, respectively. Univariable and multivariable COX regression models and Kaplan–Meier curves were used to determine the association between HER-2 expression and clinicopathological outcomes, including Gleason score (GS), pathological T (pT) stage, positive surgical margins (PSM), and BCR-free survival, respectively. Results: The median follow-up time was 43 months (IQR 35–49). Among the 44 patients, 20 (45.5%) exhibited HER-2 immuno-reactivity, including 14 (31.8%) with HER-2 1+, 6 (13.64%) with HER-2 2+, and 0 (0%) with HER-2 3+ staining. The proportion of patients with PSM was significantly lower in the HER-2 0 group than in those with HER-2 1+ or 2+ (25.0% vs. 65.0%, p = 0.008). Multivariable logistics regression models revealed that HER-2 1+ or 2+ was an independent risk factor that was strongly associated with a higher proportion of PSM (OR, 2.69; 95% CI, 0.62–11.71, p = 0.042). A total of 18 (40.9%) patients experienced BCR after surgery, including 6 (25%) in cohort-1 and 12 (60.0%) in cohort-2 (p = 0.019), as well as 13 (34.2%) in cohort-3 and 5 (83.3%) in cohort-4 (p = 0.023). Kaplan–Meier analysis showed that patients in cohort-1 (HER-2 0) had significantly longer BCR-free survival than those in cohort-2 (HER-2 1+ or 2+) (p < 0.001), and those in cohort-3 had longer BCR-free survival than those in cohort-4 (p < 0.001). Furthermore, patients with PSM showed significantly shorter BCR-free survival compared to those with patients with negative surgical margins (NSM) (p = 0.005). Multivariable Cox regression analysis revealed that HER-2 1+, 2+ (HR, 17.00; 95% CI, 1.38–210.22, p < 0.001), HER-2 2+ (HR, 2.85; 95% CI, 1.23–3.25, p = 0.004), and PSM (HR, 6.12; 95% CI, 3.08–11.72, p = 0.007) were all significant independent predictors of BCR following surgery. Conclusions: HER-2 expression is a common phenomenon in PCa; nearly half of the proportion of localized PCa had HER-2 1+ or 2+, but the cases that expressed HER-2 3+ were rare. Cases with HER-2 1+ or 2+ were more likely to develop BCR compared with HER-2 0. The HER-2 1+ or 2+ expression was closely associated with a higher incidence of PSM and was an independent predictor of shorter BCR-free survival in patients with localized prostate cancer after radical prostatectomy. Full article

Toxoplasma gondii, the causative agent of toxoplasmosis widespread in animals and humans, is an intracellular apicomplexan protozoan parasite infecting a variety of host cells. Gene editing using CRISPR-Cas9 has become a standard tool to investigate the molecular genetics of this interaction. With respect to gene knock-out (KO) studies, the general paradigm implies that the gene of interest is expressed in the wildtype and that only the gene of interest is affected by the knock-out. Consequently, the observed phenotype depends on the presence or absence of genes of interest. To challenge this paradigm, we knocked out two open reading frames (ORFs) constitutively expressed in T. gondii ShSp1 tachyzoites, but not essential, namely ORF 297720 encoding a trehalose-6-phosphatase homolog and ORF 319730 encoding a You2 C2C2 zinc finger homolog. We analyzed the proteomes of tachyzoites isolated at a late stage of infection, as well as intracellular tachyzoites and host cells at an early stage of infection. The intended KO proteins were present in the T. gondii Sp1 wildtype but absent in the KO clones. Moreover, besides differentially expressed (DE) proteins specific to each KO, 17 DE proteins common to both KOs were identified in isolated tachyzoites and 39 in intracellular tachyzoites. Moreover, 76 common DE proteins were identified in host cells. Network and enrichment analyses showed that these proteins were functionally related to antiviral defense mechanisms. These results indicate that the KO of a gene of interest may not only affect the expression of other genes of the target organism, which in our case is T. gondii, but also the gene expression of its host cells. Therefore, phenotypes of KO strains may not be causally related to the KO of a given gene. Overall, this study highlights that genetic manipulation in T. gondii can lead to system-wide proteomic shifts in both parasite and host, emphasizing the need for cautious interpretation of knock-out-based functional analyses. Full article

Background/Objectives: As crucial natural predators, hornets contribute to ecosystem function by preying on agricultural and forest pests and facilitating plant pollination. However, the predatory preference of hornets for honeybees poses a significant threat to honeybee pollination and the development of the beekeeping industry. Foraging and pollination behaviors in hornets are largely governed by a sensitive olfactory system, but their olfactory molecular mechanisms remain poorly understood. Methods: VvelCSP1 and VvelCSP4 were successfully expressed in the prokaryotic expression system and purified by Ni-NTA affinity chromatography column. Fluorescence competitive binding assays were employed to evaluate their binding affinities to volatile compounds derived from the seed elaiosome of Stemona tuberosa and honeybees. Molecular docking was further performed to analyze key residues and interaction patterns within the binding pockets. Results: Fluorescence competitive binding assays showed that both proteins prefer long-chain alkanes yet exhibit significant substrate selectivity and high ligand specificity. VvelCSP1 specifically binds to hexacosane, while VvelCSP4 specifically recognizes docosane. Molecular docking results demonstrated that the binding process between VvelCSP1, VvelCSP4 and their respective ligands is dominated by hydrophobic interactions. Conclusions: This study provides functional evidence for investigating the olfactory molecular regulation mechanisms underlying hornet-mediated seed dispersal. These findings establish a foundation for potential applications of hornets in plant propagation, biological pest control, crop pollination and ecological balance maintenance in agroforestry systems. Full article

Background: Pancreatic cancer (PC) is one of the cancers with poor prognosis. Although carbohydrate antigen (CA) 19-9 is currently the most specific and sensitive biomarker for the diagnosis of PC and, therefore, the most frequently used, approximately 34% of PC patients may have a Lewis antigen-negative phenotype and secrete little, if any, CA19-9. Therefore, effective alternative diagnostic methods for PC is required. Methods: In this study, we examined whether differentially expressed proteins in the blood of PC patients could be identified by global shotgun proteomics analysis using an in vitro cell line sample. Results: We identified 142 candidate proteins with differential expression in PC cells. A semiquantitative method and bioinformatic analysis led to a total of 14 candidate proteins that could potentially be detected in blood. Validation studies revealed that the expression of fibulin-1 was lower in PC cells than in normal pancreatic ductal cells. Moreover, in vivo fibulin-1 expression was significantly lower in serum from PC patients than in healthy individuals. Conclusions: These findings suggest that lower serum levels of fibulin-1 may be a novel biomarker for the detection of PC. Full article

Chronic alcohol misuse is the leading cause of non-ischemic dilated cardiomyopathy, and the molecular mechanisms underlying the development of alcohol-associated cardiomyopathy (ACM), particularly regarding sex-specific susceptibility and mitochondrial contributions, are not fully known. In this study, we utilized a preclinical model of chronic + binge ethanol consumption to investigate sex differences in disease severity and mitochondrial function. Male and female C57BL/6J mice were fed ethanol or control liquid diets for 30 days, with 2 binge episodes on days 10 and 30. Cardiac morphology was assessed via echocardiography and cardiac function via left ventricular pressure–volume catheterization. Mitochondrial function was evaluated ex vivo using Seahorse XF analysis, ATP luminescence, and AmplexTM Red fluorescence in isolated ventricular mitochondria. Ethanol feeding induced significant cardiac dysfunction and increased transcriptional expression of inflammatory and fibrotic markers in males, while these effects were not seen in females. Despite these sex-specific cardiac effects, mitochondrial respiration, ATP production, collagen protein expression, and oxidative stress were not significantly altered following alcohol exposure in either sex. Further investigation is warranted to assess the potential role of ovarian hormones in this female cardioprotection against chronic + binge ethanol. Full article

Systemic sclerosis (SSc) is an autoimmune disease marked by fibrosis in various organs, including the heart. Cardiac involvement influences prognosis, but the underpinning mechanisms remain unclear. This study investigates myocardial changes in a murine SSc model induced by subcutaneous injection of HOCl, with a specific focus on alterations in structural proteins and inflammatory markers, oxidative stress, and vascular remodeling. Hearts were collected from SSc mice after 6 weeks, and structural, inflammatory, and oxidative stress markers were evaluated by Hematoxylin and Eosin (H&E) staining, Masson’s Trichrome, and immunohistochemical analysis. Increased vimentin and α-SMA expression were detected in the vasculature, indicating endothelial dysfunction and myofibroblast activation, alongside a decrease in CD31 expression, consistent with an endothelial-to-mesenchymal transition (EndMT). Concomitant increases in macrophage (CD68, F4/80, EP29, EPR1) and inflammasome markers (Caspase-1, IL-1β and NLPR-3) were observed together with a remarkably augmented level of MMP3, MMP9, Collagen I and TGF-β, thus suggesting that inflammation and matrix remodeling correlate with endothelial dysfunction. Accordingly, the increased levels of NRF2 and HOMX1 suggested a compensatory protective response against oxidative stress. These data suggest that both immune cell- and inflammasome-mediated inflammation signaling play a key role in endothelial dysfunction by altering the balance between fibrosis and vascular remodeling markers. Full article

Neuromast cells are specialized mechanosensory receptor cells embedded within the lateral line system of aquatic vertebrates, enabling the detection of water movement and vibration that are essential for navigation, prey capture, and predator avoidance. These cells share common evolutionary and functional homology with mammalian inner ear hair cells, both of which rely on stereocilia-mediated mechano-transduction and ion channel activation to convert mechanical stimuli into neural signals. Unlike their mammalian counterparts, neuromast hair cells possess a regenerative capacity following damage, making the lateral line system a unique model for studying hair cell regeneration and sensory restoration. This study examines the potential of the Mexican tetra (Astyanax mexicanus) as a novel model organism for investigating ototoxicity and regeneration of neurosensory hair cells. Here, we explore the cranial and trunk lateral line neuromasts, including deep canal neuromast cells located in facial bones, such as the mandible and circumorbital bones. In the present study, juvenile surface-dwelling Mexican tetra were exposed to a 500 µM neomycin for 4 h to induce targeted hair cell damage. The samples were collected at 4-, 12-, 24-, and 72 h post-exposure. Furthermore, neuromast cell viability was assessed using [2-(4-(Dimethylamino) styryl)-N-ethylpyridinium iodide] (DASPEI). Gene expression analysis revealed a modest increase in Fibroblast Growth Factor 1 (fgf1) and Axis Inhibition Protein 2 (axin2) expression following treatment; however, these changes were not statistically significant. The SRY-box transcription factor 2 (sox2) remains constant throughout the exposure and recovery period. These findings highlighted the regenerative dynamics of neuromast cells in Mexican tetra. This work lays the foundation for future therapeutic strategies targeting human sensory deficits, particularly those involving inner ear hair cell degeneration. Full article

Obesity, a global health crisis, is linked to chronic low-grade inflammation and an increased risk of developing various chronic diseases, including psoriasis. Probiotics, postbiotics, and fermented foods have shown promise in combating inflammation and obesity. This study aimed to develop and characterize a chicory extract fermented with Bacteroides fragilis (C-B. fragilis) and its supernatant (phyto-postbiotic supernatant, PPS) as potential treatments for obesity, inflammation, and psoriasis. Polyphenols, organic acids, and amino acids were identified in the metabolic profile of C-B. fragilis. PPS and C-B. fragilis extract both revealed potent anti-inflammatory, anti-obesity, and antioxidant activities. In vitro assays highlighted that PPS significantly reduced the production of reactive oxygen species (ROS), the expression of pro-inflammatory cytokines (TNF-α, IL-6, IL-8) in macrophages, and the secretion of IL-1β in LPS-stimulated PBMCs. Moreover, PPS decreased triglyceride content in human adipocytes and modulated the expression of leptin and adiponectin. Regarding psoriasis, PPS reduced pro-inflammatory cytokines (IL-6, IL-1β) in both psoriatic keratinocytes and a co-culture model mimicking the skin-adipose tissue interface. In addition, PPS lowered S100 calcium-binding protein A7 (S100A7) expression in the co-culture model, suggesting a potential role in restoring skin barrier function. In summary, our results highlight the potential of PPS extract (supernatant of chicory fermentation by Bacteroides fragilis) as a promising therapeutic strategy for the management of obesity-related inflammation and psoriasis. Full article

Calcium/calmodulin-dependent protein kinase II (CaMKII) comprises multiple isoforms with distinct nuclear variants that exert transcriptional control in a context-dependent manner. Among them, CaMKIIδB and δ9 in the heart, and CaMKIIγ in the nervous system, have emerged as regulators of chromatin dynamics, transcription factor activity, and developmental gene programs. Nuclear localization is driven by splice-dependent nuclear localization sequences, with phosphorylation at defined serine residues modulating import and retention. Evidence supports CaMKII-dependent phosphorylation of class IIa HDACs (Ser467/Ser632 in HDAC4), linking CaMKII to MEF2 activation in cardiac hypertrophy, and interactions with NF-κB and HSF1 further expand its nuclear repertoire. In the nervous system, CaMKIIγ contributes to kinase-dependent gene expression, potentially influencing plasticity and disease susceptibility. While these mechanisms highlight nuclear CaMKII as an isoform-specific regulator of transcription, direct evidence remains elusive, and several CaMKII putative substrates require further validation. This review synthesizes current knowledge on nuclear CaMKII isoforms, emphasizes established mechanistic pathways, and outlines unsolved questions critical for understanding their roles in development, disease progression, and therapeutic targeting. Full article

Endometrial carcinoma is the most frequent gynecologic malignancy in western countries. In recent years, mutations in CTNNB1 have been associated with worse prognosis in low-risk carcinomas. However, there is a lack of understanding of the proteomic implications of CTNNB1 mutations in this type of tumor. In this study, we performed shotgun proteomics using Formalin-Fixed Paraffin-Embedded (FFPE) tissue samples of CTNNB1 mutated and wild-type low-risk endometrial carcinomas. A publicly available proteomic and transcriptomic database was used to validate results. Differential protein expression and Gene Set Enrichment Analysis revealed dysregulation of pathways associated with cell keratinization, immune response modulation, and intracellular calcium regulation. CTNNB1 mutated tumors showed immune dysregulation at multiple levels including cytokine secretion, cell adhesion, and lymphocyte activation. These results were supported by tissue multiplex immunofluorescence analysis, demonstrating reduced CD8 tumor-infiltrating lymphocytes and different immune spatial interaction patterns. Intracellular calcium dysfunction was associated with key transcript dysregulation. We found an increased expression of CAMK2A and ROR2, suggesting a potential role for non-canonical Wnt pathway activation in CTNNB1 mutated tumors. Full article

Heat shock protein 70 (HSP70) represents a promising target for cancer therapy. Oncolytic vaccinia virus (oncoVV) mediates tumor regression through direct oncolysis and immune activation. However, the anti-tumor potential of HSP70-silenced oncoVV (oncoVV-shHSP70) remains unexplored. Here, we demonstrate that oncoVV-shHSP70 achieves superior tumor regression in ovarian cancer models (cell lines, immunodeficient mice and humanized mice) via dual mechanisms including enhancing apoptosis, autophagy flux, ROS generation, and immune reprogramming. Notably, we found that oncoVV-shHSP70 triggers an autophagy–ROS feedback loop that amplifies viral replication and pro-inflammatory cytokine expression. Crucially, in humanized mice, oncoVV-shHSP70 induced spatial redistribution of cytotoxic T cells, expanding tumor-infiltrating hCD8⁺hGZMB⁺ populations. These findings position oncoVV-shHSP70 as a promising viro-immunotherapy that co-opts HSP70 silencing to potentiate both direct oncolysis and anti-tumor immunity, providing a preclinical rationale for viro-immunotherapy in solid tumors. Full article

Global temperature rise has become a critical challenge to agricultural sustainability, severely affecting crop growth, productivity, and survival. Human-induced climate change and greenhouse gas emissions cause heat stress, disrupting plant metabolism and physiology at all developmental stages from germination to harvest. Elevated temperatures during germination impair water uptake, enzyme activity, and energy metabolism, leading to poor or uneven seedling emergence. At key phases such as flowering and grain filling, heat stress limits photosynthesis and transpiration by inducing stomatal closure, restricting carbon dioxide intake, and reducing photosynthetic efficiency. The reproductive stage is particularly vulnerable to high temperatures, impairing pollen viability, preventing anther dehiscence, and reducing fertilization success. Membrane instability further accelerates chlorophyll degradation and leaf senescence. Heat stress also alters biochemical and hormonal balances by disrupting the synthesis and signaling of auxins, gibberellins, and abscisic acid (ABA). Elevated ABA promotes stomatal closure to enhance stress tolerance, while increased ethylene levels trigger premature leaf senescence and abscission. These hormonal shifts and oxidative stress hinder plant growth and reproduction, threatening global food security. Although plants employ adaptive mechanisms such as heat shock protein expression and stress-responsive gene regulation, current strategies remain inadequate, highlighting the urgent need for innovative approaches to improve crop resilience under rising temperatures. Full article